Many clinical labs rely on uracil DNA glycosylase (UDG) (also known as uracil N-glycosylase (UNG)) decontamination of polymerase chain reaction (PCR) products. Amplicons containing uracil as opposed to thymidine can be digested with UDG to eliminate any residual PCR product in the laboratory. Many Next Generation sequencing platforms utilize polymerases that cannot traverse a uracil (utilize polymerases that are uracil illiterate). For instance, the Illumina MiSeq and HiSeq platforms rely on polymerases with proof reading activity to generate the seeded clusters for surface PCR. Proof reading polymerases such as pfu will stall on uracils in the template strand. Due to this, amplification of uracilyated templates fail to initiate cluster PCR thus eliminating the potential of UDG decontamination methods.
Thus, improved amplification methods and/or decontamination of amplification methods are needed for nucleic acid amplification techniques such as PCR.